Saleh Ola A, El-Azzouny Aida A, Aboul-Enein Hassan Y, Badawey Amr M
Medicinal and Pharmaceutical Chemistry Department, Pharmaceutical and Drug Industries Research Division, National Research Centre, Dokki-Giza, Egypt.
Medicinal and Pharmaceutical Chemistry Department, Pharmaceutical and Drug Industries Research Division, National Research Centre, Dokki-Giza, Egypt
J Chromatogr Sci. 2014 Jul;52(6):539-46. doi: 10.1093/chromsci/bmt062. Epub 2013 Jun 5.
A stability-indicating reversed-phase high-performance liquid chromatographic assay procedure has been developed and validated for riluzole in the presence of alkaline and oxidative degradation products. The liquid chromatographic separation was achieved and compared isocratically on C18 Zorbax ODS and Poroshell 120 EC-C18 columns by using a mobile phase containing methanol-water, pH = 3.10 (70:30, v/v), at a flow rate of 1 mL/min and ultraviolet detection at 264 nm. The method was linear over the concentration ranges of 20-200 µg/mL (r = 0.9997) and 10-200 µg/mL (r = 0.9995). The limit of detection and quantitation for the two columns were 2 and 6 µg/mL and 1 and 3 µg/mL, respectively. Moreover, spectrophotometric methods were applied for the determination of riluzole in the presence of its oxidative degradation products by using first derivative spectrophotometry at 252.5 and 275.0 nm. The method was linear over the concentration range of 1-20 µg/mL (r = 0.9995 and 0.9996) at the studied wavelengths, with limits of detection and quantitation of 0.1 and 0.3 µg/mL. In addition, the first derivative ratio spectrophotometry (DD1) method was applied for the determination of riluzole in the presence of its alkaline degradation product at 252.0, 278.5 and 306.3 nm by using 100 µg/mL of alkaline degraded riluzole as a divisor; riluzole was additionally determined in the presence of its hydrogen peroxide oxidative degradation products at 252.5, 275.0 and 305.0 nm by using 200 µg/mL of oxidative degraded riluzole as a divisor. The DD1 method was linear over the concentration range of 1-20 µg/mL (r = 0.9996, 0.9995 and 0.9996 for the alkaline degradation product at the three studied wavelengths, respectively; and r = 0.9995, 0.9996 and 0.9995 for the oxidative degradation product at the three studied wavelengths, respectively), with limits of detection and quantitation of 0.1 and 0.3 µg/mL for both alkaline and oxidative degradation products. The two studied chromatographic and spectrophotometric methods were comparable and display the required accuracy, selectivity, sensitivity and precision to assay riluzole in bulk and pharmaceutical dosage forms. Degradation products resulting from the stress studies did not interfere with the detection of riluzole, which indicates that these are stability-indicating assays.
已开发并验证了一种稳定性指示反相高效液相色谱分析方法，用于测定在碱性和氧化降解产物存在下的利鲁唑。在C18 Zorbax ODS和Poroshell 120 EC-C18柱上，以甲醇 - 水（pH = 3.10，70:30，v/v）为流动相，流速为1 mL/min，在264 nm处进行紫外检测，实现了液相色谱分离并进行了等度比较。该方法在20 - 200 µg/mL（r = 0.9997）和10 - 200 µg/mL（r = 0.9995）的浓度范围内呈线性。两根色谱柱的检测限和定量限分别为2和6 µg/mL以及1和3 µg/mL。此外，采用一阶导数分光光度法，在252.5和275.0 nm波长下，对存在氧化降解产物的利鲁唑进行分光光度法测定。该方法在所研究的波长下，在1 - 20 µg/mL的浓度范围内呈线性（r = 0.9995和0.9996），检测限和定量限分别为0.1和0.3 µg/mL。另外，采用一阶导数比率分光光度法（DD1），以100 µg/mL碱性降解的利鲁唑为除数，在252.0、278.5和306.3 nm波长下测定存在碱性降解产物时的利鲁唑；以200 µg/mL氧化降解的利鲁唑为除数，在252.5、275.0和305.0 nm波长下测定存在过氧化氢氧化降解产物时的利鲁唑。DD1方法在1 - 20 µg/mL的浓度范围内呈线性（对于碱性降解产物，在三个研究波长下r分别为0.9996、0.9995和0.9996；对于氧化降解产物，在三个研究波长下r分别为0.9995、0.9996和0.9995），碱性和氧化降解产物的检测限和定量限均为0.1和0.3 µg/mL。所研究的两种色谱和分光光度法具有可比性，并且在测定原料药和药物剂型中的利鲁唑时，显示出所需的准确性（准确度）、选择性、灵敏度和精密度。加速试验产生的降解产物不干扰利鲁唑的检测，这表明这些是稳定性指示分析方法。
